Skip to main content
STI & HIV 2023 World Congress Main banner
Final Program


Icon Legend
Back

Clinical Science

Category: Laboratory methods

(P191) Validation of pooling triple site samples to detect STIs using the Sacace Neisseria gonorrhoeae/Chlamydia trachomatis/Mycoplasma genitalium/Trichomonas vaginalis Real-TM kit.

Tuesday, July 25, 2023
16:00 – 18:00 CST
Background: Extra-genital Sexually transmitted infections (STIs) are mostly asymptomatic and if not detected, may facilitate onward transmission. However, testing three biological sites (urethra, pharynx, and anorectum) is expensive but pooling strategies whereby samples of the three anatomical sites are combined in one sample, showed to be cost-effective. Validated pooling strategies mostly use large testing platforms which are difficult to access in Cuba. The aim of this work was to validate the SacaceTM
N. gonorrhoeae/С. trachomatis/M. genitalium/T. vaginalis Real-TM kit which can be used on the Rotor-Gene Q instrument which is available in Cuba.



Methods: Left-over urine, pharyngeal, and anorectal swab samples of patients that came to our clinic for STI testing and were positive for either N. gonorrhoeae, С. trachomatis and M. genitalium with high and low bacterial load were analyzed. A total of 500 µL of diluted PBS was added to the swab. A panel of eight pools (see Table 1) was created by combining 100 μL of each sample type to a final volume of 300 uL. Automated DNA extraction was performed using the QIAamp DNA Mini Kit (QIAagen, Germany) and the QIAcube automated station (QIAgen). PCR amplification of the pooled and individual samples was performed in a Rotor-Gene Q instrument in the same run.

Results: Results are shown in Table 1. All results of the pool were in concordance with the final result of the individual samples. However, in most of the cases, an increase in cycle threshold values of the pooled sample was observed, which may indicate a lower bacterial load in the pooled sample, probably due to dilution.



Conclusion: Although this study was limited in sample size, these data may show that the Sacace kit might be used in a pooled way to detect STIs in genital and extra-genital sites. As such, extra-genital screening of CT/NG using this pooled strategy may be implemented in Cuba. Although the need to test for MG is discussed, the pooling methodology using the Sacace kit may be used in future surveillance studies.